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Identification of Mammalian Mediator Subunits with Similarities to Yeast Mediator Subunits Srb5, Srb6, Med11, and Rox3

Sato, Shigeo and Tomomori-Sato, Chieri and Banks, Charles A. S. and Sorokina, Irina and Parmely, Tari J. and Kong, Stephanie E. and Jin, Jingji and Cai, Yong and Lane, William S. and Brower, Christopher S. and Conaway, Ronald C. and Conaway, Joan Weliky (2003) Identification of Mammalian Mediator Subunits with Similarities to Yeast Mediator Subunits Srb5, Srb6, Med11, and Rox3. Journal of Biological Chemistry, 278 (17). pp. 15123-15127. ISSN 0021-9258. http://resolver.caltech.edu/CaltechAUTHORS:SATjbc03b

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Abstract

The Mediator is a multiprotein coactivator required for activation of RNA polymerase II transcription by DNA binding transactivators. We recently identified a mammalian homologue of yeast Mediator subunit Med8 and partially purified a Med8-containing Mediator complex from rat liver nuclei (Brower, C. S., Sato, S., Tomomori-Sato, C., Kamura, T., Pause, A., Stearman, R., Klausner, R. D., Malik, S., Lane, W. S., Sorokina, I., Roeder, R. G., Conaway, J. W., and Conaway, R. C. (2002) Proc. Natl. Acad. Sci. U. S. A. 99, 10353-10358). Analysis of proteins present in the most highly purified Med8-containing fractions by tandem mass spectrometry led to the identification of many known mammalian Mediator subunits, as well as four potential Mediator subunits exhibiting sequence similarity to yeast Mediator subunits Srb5, Srb6, Med11, and Rox3. Here we present direct biochemical evidence that these four proteins are bona fide mammalian Mediator subunits. In addition, we identify direct pairwise binding partners of these proteins among the known mammalian Mediator subunits. Taken together, our findings identify a collection of novel mammalian Mediator subunits and shed new light on the underlying architecture of the mammalian Mediator complex.


Item Type:Article
Additional Information:Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc. Received for publication, February 5, 2003. Originally published In Press as doi:10.1074/jbc.C300054200 on February 12, 2003 We thank R. Roeder and S. Malik for the HeLa cell line expressing FLAG-Nut2. This work was supported by National Institutes of Health Grant R37 GM41628.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. The on-line version of this article (available at http://www.jbc.org) includes as Supplemental Material (i) a sequence comparison of yeast and mammalian Mediator subunits and (ii) peptide sequences obtained from the purified rat liver Mediator complex.
Record Number:CaltechAUTHORS:SATjbc03b
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:SATjbc03b
Alternative URL:http://dx.doi.org/10.1074/jbc.C300054200
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:6958
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:03 Jan 2007
Last Modified:26 Dec 2012 09:27

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